Medium development for production of Bacillus thuringiensis based biopesticides


Tezin Türü: Yüksek Lisans

Tezin Yürütüldüğü Kurum: Orta Doğu Teknik Üniversitesi, Fen Edebiyat Fakültesi, Biyolojik Bilimler Bölümü, Türkiye

Tezin Onay Tarihi: 2008

Öğrenci: ORHAN ÖZCAN

Danışman: GÜLAY ÖZCENGİZ

Özet:

The insect pathogen Bacillus thuringiensis (Bt) holds great promise as an effective and friendly way for management of the pests with safety for nontarget animals and humans. However, high capital investment due to high production and formulation cost of commercial Bt preparations has caused prohibitive effect on companies. The present study mainly aimed at developing a low cost medium that supports the growth of different Bt strains and their specific bioinsecticidal δ-endotoxins (crystal proteins). A comparison was made between the representative members of three different subspecies of Bt to observe toxin yields in response to certain nutritional conditions. Three different Bt subspecies were Bt kurstaki (strain 81), Bt israelensis (strain HD500) and Bt tenebrionis (strain 3203), producing lepidoptera- and diptera-specific Cry1 and Cry2, diptera-specific Cry4Ba and Cry11Aa and coleoptera-specific Cry3Aa toxins, respectively. Studies were conducted to optimize glucose and inorganic phosphate concentrations in standard DSM medium for the production of these Bt-based biopesticides. General suppression of toxin yields in high glucose medium (10 g/L) thought the generality of carbon catabolite regulation for biosynthesis of different types of toxins. Inorganic phosphate (Pi) level was important for Cry4Ba, Cry11Aa and Cry3Aa biosynthesis while Cry1 and Cry2 production was not responsive to high Pi. Wastewater sludge, fruit residues and broiler litter were next tested as cheap raw materials for Bt-based biopesticide production in batch cultures. Broiler litter seemed to be a much better substrate among all since some degree of production of each toxin was observed at almost every stage of fermentation. The processing of broiler litter was found to significantly improve toxin yields. The medium prepared from processed broiler litter was successfully used to cultivate all Bt stains and obtain bioinsecticidal proteins in high yields which were comparable or higher than those that can be obtained on standard semi-synthetic media.